Figure 9.

Cytokines and α-synuclein expression. (A) We tested cytokines to determine if they could induce increased expression of α-syn in SH-SY5Y cells. SH-SY5Y cells were grown in serum free medium and 50 ng/mL of either mouse TNFα, IL-1β, or both was applied to the cells twice in a 24 h period. Western blot was then carried out to assess α-syn expression levels. Tubulin was also assessed as a loading control. (B) Analysis showed that only TNFα or TNFα/IL-1β caused an increase in α-syn expression. As IL-1β had no significant effect this result was only due to the presence of TNFα. (C) We also tested the effects of the cytokines on the activity of the SNCA promoter reporter −6.1/−1.3 which showed the strongest response to iron-fed conditioned medium. The SH-SY5Y cells transiently transfected with the reporter were treated similarly with the cytokines. Luciferase activity was significantly (p < 0.05) increased only for cells treated with TNFα alone or in combination with IL-1β. Shown are the mean and S.E.M. of four separate experiments.